<div class="csl-bib-body">
<div class="csl-entry">Schittmayer, M., Mitulovic, G., Krawitzky, M., Kruppa, G., & Birner-Grünberger, R. (2024, June 2). <i>nano-lipidomics employing silica based monolithic columns</i> [Poster Presentation]. 72nd ASMS Conference 2024, Anaheim, United States of America (the). http://hdl.handle.net/20.500.12708/207568</div>
</div>
-
dc.identifier.uri
http://hdl.handle.net/20.500.12708/207568
-
dc.description.abstract
Nano-liquid chromatography is the state of the art in the proteomics field, yet lipidomics applications that make use of the vastly increased efficiency of electrospray ionization at sub-µL flow rates are still considered exotic. A handful of recent studies have shown that additional experimental efforts that are commonly associated with chromtaographic downsizing are vastly outmatched by the gain of sensitivity and concomitant achieved lipid species coverage. Considering the dynamic concentration range of the lipidome which is spanning from the pico- to the millimolar range, it is obvious that analysis of low abundant lipid will greatly benefit from improved sensitivity. Also, single-cell lipidomics is an upcoming application that will greatly benefit from employing nanoscale liquid chromatography.
Nano-lipidomics analysis on a novel silica monolithic column (C18, 75 µm, 150 mm length) was compared to a conventional microbore setup (ACQUITY UPLC BEH C18 Column, 130Å, 1.7 µm, 1 mm X 150 mm). A synthetic standard mixture (791500, Avanti Lipids) was diluted in a folch extract of human squamous cell carcinoma cell line SAS. Total amounts for individual lipid species ranged from 192 pmol to 3.3 fmol on column. Samples were analyzed in both polarities utilizing either the Bruker VIP-HESI source or the captive spray source employing the 4D-lipidomics PASEF method. Spiked standard lipids were subjecte4d to targeted analysis employing Skyline 23.1.0.380 and background lipidomes were analyzed in an untargeted manner employing Metaboscape 2023.
Equal amounts of each standard dilution was injected in 4 technical replicates on both front ends from identical vials. With the exception of lysoPC, which was not retained on the nano-lipidomics setup without further optimization, both setups showed comparable chromatographic performance and quantitative accuracy in the upper concentration range. As expected, peak areas were on average ~ 200fold higher on the nano-lipidomics setup and hence data quality at low concentrations was significantly improved. For example, at the lowest tested concentration (femtomol range OC) for most lipid species M+2 was at or below the LOD on the microbore setup, leading to a dropoff of idotp score while the nano-lipidomics setup yielded consistent values > 0.9. The most pronounced effect was observed for PI 18:1 where even the monoisotopic peak was below LOD on the microbore setup whereas in the nanolipidomic setup the M+2 peak was readily detectable and the overal isotopic pattern was scored at 0.95. Untargeted analysis employing basic rule based annotation yielded 242 species (158 positive and 84 negative mode) on the microbore setup. The identical sample load on the nano-lipidomics setup yielded almost double the number with 506 species (332 positive and 172 negative mode), respectively.
Nanoflow lipidomics employing monolithic columns allows identification quantification of low abundant lipids outperforming a state of the art microbore setup.
en
dc.language.iso
en
-
dc.subject
nano-lipidomics
en
dc.subject
chromatography column
en
dc.title
nano-lipidomics employing silica based monolithic columns
en
dc.type
Presentation
en
dc.type
Vortrag
de
dc.contributor.affiliation
Bruker (Switzerland), Switzerland
-
dc.contributor.affiliation
Bruker Corporation (Billerica, US)
-
dc.contributor.affiliation
Life Science Mass Spectrometry - Bruker SRO (Brno, CZ)
-
dc.type.category
Poster Presentation
-
tuw.researchTopic.id
M2
-
tuw.researchTopic.name
Materials Characterization
-
tuw.researchTopic.value
100
-
tuw.publication.orgunit
E164-01-3 - Forschungsgruppe Bioanalytik
-
tuw.author.orcid
0000-0003-3249-655X
-
tuw.author.orcid
0000-0002-5906-5816
-
tuw.author.orcid
0000-0002-4844-9865
-
tuw.author.orcid
0000-0003-3950-0312
-
tuw.event.name
72nd ASMS Conference 2024
en
tuw.event.startdate
02-06-2024
-
tuw.event.enddate
06-06-2024
-
tuw.event.online
On Site
-
tuw.event.type
Event for scientific audience
-
tuw.event.place
Anaheim
-
tuw.event.country
US
-
tuw.event.institution
ASMS
-
tuw.event.presenter
Schittmayer, Matthias
-
wb.sciencebranch
Chemie
-
wb.sciencebranch.oefos
1040
-
wb.sciencebranch.value
100
-
item.languageiso639-1
en
-
item.openairetype
conference poster not in proceedings
-
item.grantfulltext
none
-
item.fulltext
no Fulltext
-
item.cerifentitytype
Publications
-
item.openairecristype
http://purl.org/coar/resource_type/c_18co
-
crisitem.author.dept
E164-01-3 - Forschungsgruppe Bioanalytik
-
crisitem.author.dept
Bruker (Switzerland)
-
crisitem.author.dept
Bruker Corporation (Billerica, US)
-
crisitem.author.dept
Life Science Mass Spectrometry - Bruker SRO (Brno, CZ)
-
crisitem.author.dept
E164-01 - Forschungsbereich Imaging und Instrumentelle Analytische Chemie
-
crisitem.author.orcid
0000-0003-3249-655X
-
crisitem.author.orcid
0000-0002-5906-5816
-
crisitem.author.orcid
0000-0002-4844-9865
-
crisitem.author.orcid
0000-0003-3950-0312
-
crisitem.author.parentorg
E164-01 - Forschungsbereich Imaging und Instrumentelle Analytische Chemie
-
crisitem.author.parentorg
E164 - Institut für Chemische Technologien und Analytik
